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Melanocytes in vitro: how do they undergo mitosis?
S Kippenberger1, A Bernd, J Bereiter-Hahn
1Zentrum der Dermatologie und Venerologie, Klinikum der Johann Wolfgang Goethe Universität, Frankfurt/Main, Germany.
Pigment Cell Research
|February 1, 1997
Summary
Human skin melanocytes, typically slow-cycling, undergo significant morphologic changes during mitosis in vitro. Centrifuging spent medium and re-adding detached mitotic cells enhances melanocyte cultures.
Area of Science:
- Dermatology
- Cell Biology
- In Vitro Cell Culture
Background:
- Human adult skin melanocytes are characterized by slow cycling and a dendritic morphology.
- In vitro proliferation of melanocytes can be induced with appropriate stimulators.
- Melanocyte mitosis involves distinct morphologic alterations, including cell rounding and detachment.
Purpose of the Study:
- To investigate the morphologic changes of human melanocytes during mitosis in vitro.
- To identify optimal methods for maintaining and enhancing melanocyte proliferation in culture.
Main Methods:
- Time-lapse microscopy was employed to observe melanocyte behavior during cell division.
- Quantification of detached, viable, and mitotic cells was performed.
- Experimental manipulation involving medium changes and centrifugation was conducted.
Main Results:
- Melanocyte mitosis involves retraction of dendrites, cell rounding, detachment from the substrate, and suspension during division.
- Approximately 1% of cells in culture are detached, with about 70% of these being viable and in mitosis.
- Standard medium changes lead to the loss of mitotic cells, favoring quiescent (G0) cells.
Conclusions:
- Melanocyte division occurs in suspension after retracting dendrites and rounding.
- Centrifugation of exhausted medium and reintroduction of detached mitotic cells is recommended to improve melanocyte culture yield.
- This technique supports the selection and retention of actively dividing melanocytes.